Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-09-23T14:40:10Z No. of bitstreams: 1 luisamariasilveiradealmeida.pdf: 3706782 bytes, checksum: 1b4e01bcd663e5b0167c387beb0d0e2d (MD5) / Approved for entry into archive by Diamantino Mayra (mayra.diamantino@ufjf.edu.br) on 2016-09-26T20:31:05Z (GMT) No. of bitstreams: 1 luisamariasilveiradealmeida.pdf: 3706782 bytes, checksum: 1b4e01bcd663e5b0167c387beb0d0e2d (MD5) / Made available in DSpace on 2016-09-26T20:31:05Z (GMT). No. of bitstreams: 1 luisamariasilveiradealmeida.pdf: 3706782 bytes, checksum: 1b4e01bcd663e5b0167c387beb0d0e2d (MD5) Previous issue date: 2015-07-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A esquistossomose é uma doença que afeta cerca de 200 milhões de pessoas no mundo e mais de 600 milhões estão expostas ao risco de contraí-la, sendo que no Brasil, o Estado de Minas Gerais é um dos mais afetados com esta parasitose. Atualmente, o único fármaco disponível para o tratamento desta doença é o praziquantel (PZQ). No entanto, este medicamento já está no mercado há décadas e tem demonstrado relatos de ineficácia. Neste contexto, extratos e metabólitos secundários obtidos de espécie do gênero Artemisia (Asteraceae) têm demonstrado expressiva atividade esquistossomicida. Assim, o presente trabalho descreve o estudo fitoquímico do extrato diclorometânico das folhas de Artemisia absinthium L. (AAF), bem como a avaliação da atividade esquistossomicida in vitro dos extratos AAF e das raízes, do óleo essencial e do metabólito obtido. O estudo fitoquímico, do AAF, culminou no isolamento e identificação de dois metabólitos: uma flavona, artemetina e uma lactona sesquiterpênica, hidroxipelenolídeo. No estudo esquistossomicida in vitro foi utilizado vermes adultos de S. mansoni da linhagem BH e os parâmetros avaliados foram mortalidade, motilidade e alterações tegumentares. Os resultados deste estudo demonstraram que o extrato diclorometânico bruto (AAF), na concentração de 200 μg/mL, possui expressiva atividade esquistossomicida in vitro ao ser capaz de causar a morte de 100% dos parasitos após 24 h de incubação. Em relação à atividade esquistossomicida do extrato bruto da raiz (200 μg/mL), este foi capaz de provocar a morte de todos os vermes após 48 h. O óleo essencial da A. absinthium L. (100 µg/mL) apresentou atividade esquistossomicida moderada sendo capaz de causar 60% de mortalidade nos parasitos após 48 h de incubação. Dentre as substâncias isoladas, a artemetina, foi submetida ao ensaio de atividade esquistossomicida e não foi ativa. Estudos adicionais de avaliação esquistossomicida devem ser realizados com o extrato bruto e metabólitos de A. absinthium L. com o intuito de se identificar as moléculas responsáveis pela atividade esquistossomicida in vitro observada pelo extrato AAF. / Schistosomiasis is a disease that affects more than 200 million individuals in the world and 600 million people remain under risk of infection. In Brazil, the Minas Gerais state is one of the most affected by this disease. Nowadays, the only drug available for schistosomiasis treatment is praziquantel (PZQ). However, this drug is already on market for decades and has shown ineffectiveness. In this context, extract and secondary metabolites of the genus Artemisia (Asteraceae) have shown an expressive schistosomicidal activity. Thus, the present work describes the phytochemical study of the dichloromethane extract of Artemisia absinthium L. leaves (AAF), as well as in vitro schistosomicidal evaluation of AAF, crude roots extracts, essential oil and metabolite. The phytochemical study of AAF allowed the isolation and identification of two metabolites: one flavone, artemetin and one sesquiterpene lactone, hydroxypelenolide. In the in vitro schistosomicidal study, was used adult worms of the S. mansoni BH strain and the parameters evaluated were mortality, motility and alterations tegumental surface. The results showed that AAF, at 200 μg/mL, presents expressive schistosomicidal activity being able to cause 100% of death in parasites after 24 h of incubation. Regarding the schistosomicidal activity of the extract crude roots extract (200 μg/mL), was able to kill all the parasites after 48 h. The A. absinthium L. essential oil (100 mg / mL) showed moderate schistosomicidal activity being able to cause 60% mortality of parasites after 48 h of incubation. Among the isolated compounds, artemetin, was subjected to the schistosomicidal activity test and was inactive. Additional schistosomicidal assays should be carried out with A. absinthium L. crude extract and metabolites in order to identify the molecules responsible for schistosomicidal in vitro activity observed by AAF.

Malaria är en farlig tropiksjukdom orsakad av parasiten Plasmodium som vållar många dödsfall varje år. Sedan några år tillbaka rekommenderar Världshälsoorganisationen (WHO) användandet av artemisinin och dess derivat för behandlandet av malaria. Artemisinin syntetiseras normalt i växten Artemisia annua i lågt utbyte. På grund av det låga utbytet är läkemedlet väldigt dyrt. Då parasiten blivit resistent mot de flesta malarialäkemedel är artemisinin ett viktigt preparat i kampen mot malaria. Forskning pågår för att hitta nya eller effektivare metoder för framställning av substansen då en oro finns att produktionen från A. annua inte kommer kunna möta kraven från läkemedelsindustrin. En av teorierna är ifall andra växter inom Artemisia-släktet kan syntetisera artemisinin då flera växter uppvisat helande effekter vid andra sjukdomar. I denna studie undersöktes det ifall växterna A. vulgaris och A. absinthium från Artemisia-släktet skulle kunna syntetisera artemisinin. Med hjälp av molekylärbiologiska tekniker isolerades genetiskt material ifrån växterna. Materialet granskades efter ribonukleinsyra (RNA)- och deoxiribonukleinsyra (DNA) -sekvenser för funktionella enzym som katalyserar reaktioner i artemisinins biosyntes. Ifall generna uttrycks för dessa enzym kan eventuellt artemisinin bildas. Växterna hämtades från Revsudden, Sverige och genetiskt material isolerades. Förekomsten av genuttryck för fem viktiga enzym i artemisinins biosyntes undersöktes med Polymerase Chain Reaction (PCR). Resultatet blev att växterna hade genuttryck för två respektive tre av de fem enzymen. Detta pekar mot att varken A. vulgaris eller A. absinthium kan syntetisera artemisinin då de saknade några viktiga nyckelenzym i syntesen. Trots att en tidigare studie indikerar närvaro av artemisinin i dessa växter kan slutsatsen dras att A. vulgaris och A. absinthium inte kan bilda artemisinin. / Malaria is a tropical disease that accounts for the death of many people annually and is caused by a parasite called Plasmodium. The World Health Organization (WHO) recommends artemisinin and its derivates for malaria treatment. Artemisinin is synthesized generally in Artemisia annua in small amounts. The artemisinin-treatment is very expensive due to the small amounts produced in the plant. Since the parasite has developed resistance towards many antimalarial drugs, artemisinin is an important drug against malaria. Research to find alternative methods for artemisinin-production has begun because there is a great concern that artemisinin-production at current rate will not meet the demand from the pharmaceutical industry. Some speculate if artemisinin can be synthesized in other plants from the Artemisia-genus since many plants have shown healing properties towards other diseases. In this study, we investigated if A. vulgaris and A. absinthium could produce artemisinin. Using molecular biology techniques, genetic material was isolated from the plants. Ribonucleotide (RNA)- and deoxyribonucleotide (DNA)- sequences which encode important enzymes in the artemisinin biosynthesis were examined. In case all the genes were expressed, artemisinin may be synthesized. The plants were picked on Revsudden, Sweden and genetic material was isolated. The presence of gene expression of five important enzymes in the artemisinin biosynthesis was investigated by Polymerase Chain Reaction (PCR). The results showed that the plants had gene expression of two respectively three of the five enzymes. Due to the fact that the plants need all five enzymes to synthesize artemisinin, even though a recent study has shown presence of artemisinin in these plants, this study concludes that artemisinin cannot be synthesized in A. vulgaris and A. absinthium.